miR‑196a‑5p modulates gastric cancer stem cell characteristics by targeting Smad4

Pan,Yunzhi; Shu,Xiong; Sun,Lichao; Yu,Long; Sun,Lixin; Yang,Zhihua; Ran,Yuliang

doi:10.3892/ijo.2017.3965

miR‑196a‑5p modulates gastric cancer stem cell characteristics by targeting Smad4

Authors:
- Yunzhi Pan
- Xiong Shu
- Lichao Sun
- Long Yu
- Lixin Sun
- Zhihua Yang
- Yuliang Ran
View Affiliations

Affiliations: National Cancer Center/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Chaoyang, Beijing 100021, P.R. China
Published online on: April 19, 2017 https://doi.org/10.3892/ijo.2017.3965
Pages: 1965-1976
Copyright: © Pan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Cancer stem cells (CSCs) are undifferentiated cancer cells with a high tumorigenic activity, the ability to undergo self-renewal, and a multilineage differentiation potential. Clinical evidence suggests that CSCs in a tumor mass are the cellular determinants to promote cancer invasion and metastasis. MicroRNAs (miRNAs) have emerged as important modulators of cancer stem cell characteristics. Unveiling the candidate miRNAs that regulate CSCs may provide novel therapeutic targets against cancer. We analyzed the miRNA expression profiles regulating the cancer stem-like cell characteristics in gastric cancer. Gastric cancer stem cells (GCSCs) were sorted using the stem cell marker CD44 by fluorescence-activated cell sorting. Functional studies revealed that CD44(+) cells formed more sphere colonies and showed higher invasiveness than CD44(-) cells. miRNA microarray analysis revealed that miR‑196a‑5p was significantly upregulated in CD44(+) cells than CD44(-) cells. Suppression of miR‑196a‑5p led to decreased colony formation and invasion of GCSCs. miR‑196a‑5p decreased the expression of Smad4 by targeting 3'-UTR of the mRNA. The expression of Smad4 in gastric cancer tissues was correlated with differentiation state of tumors, TNM stage and depth of invasion. The stimulation of epithelial-mesenchymal transition (EMT) by miR‑196a‑5p in cancer stem-like cells was abolished by overexpression of Smad4. Collectively, these data demonstrate that miR‑196a‑5p has a key role in EMT and invasion by targeting Smad4 in GCSCs. miR‑196a‑5p may serve as a potential target for gastric cancer therapy.

View Figures

View References

1	Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J and Jemal A: Global cancer statistics, 2012. CA Cancer J Clin. 65:87–108. 2015. View Article : Google Scholar : PubMed/NCBI
2	Chen W, Zheng R, Zhang S, Zhang S, Zhao P, Li G, Wu L and He J: Report of incidence and mortality in China cancer registries, 2009. Chin J Cancer Res. 25:10–21. 2013.PubMed/NCBI
3	Kim TH and Shivdasani RA: Stomach development, stem cells and disease. Development. 143:554–565. 2016. View Article : Google Scholar : PubMed/NCBI
4	Jordan CT, Guzman ML and Noble M: Cancer stem cells. N Engl J Med. 355:1253–1261. 2006. View Article : Google Scholar : PubMed/NCBI
5	Zhang X, Hua R, Wang X, Huang M, Gan L, Wu Z, Zhang J, Wang H, Cheng Y and Li J: Identification of stem-like cells and clinical significance of candidate stem cell markers in gastric cancer. Oncotarget. 7:9815–9831. 2016.PubMed/NCBI
6	Alison MR, Lin WR, Lim SM and Nicholson LJ: Cancer stem cells: In the line of fire. Cancer Treat Rev. 38:589–598. 2012. View Article : Google Scholar : PubMed/NCBI
7	Liu TJ, Sun BC, Zhao XL, Zhao XM, Sun T, Gu Q, Yao Z, Dong XY, Zhao N and Liu N: CD133⁺ cells with cancer stem cell characteristics associates with vasculogenic mimicry in triple-negative breast cancer. Oncogene. 32:544–553. 2013. View Article : Google Scholar
8	Sakakini N, Turchi L, Bergon A, Holota H, Rekima S, Lopez F, Paquis P, Almairac F, Fontaine D, Baeza-Kallee N, et al: A positive feed-forward loop associating EGR1 and PDGFA promotes proliferation and self-renewal in glioblastoma stem cells. J Biol Chem. 291:10684–10699. 2016. View Article : Google Scholar : PubMed/NCBI
9	Nataraj SM, Prema CL, Vimalambike MG, Shivalingaiah SC, Sundaram S, Kumar AP, Math AK and Prashant A: Major protein of carcinoembryonic antigen gene family - CD66c, a novel marker in colon carcinoma. J Clin Diagn Res. 10:XC01–XC04. 2016.PubMed/NCBI
10	Liu J, Ma L, Xu J, Liu C, Zhang J, Liu J, Chen R and Zhou Y: Spheroid body-forming cells in the human gastric cancer cell line MKN-45 possess cancer stem cell properties. Int J Oncol. 42:453–459. 2013.
11	Takebe N, Harris PJ, Warren RQ and Ivy SP: Targeting cancer stem cells by inhibiting Wnt, Notch, and Hedgehog pathways. Nat Rev Clin Oncol. 8:97–106. 2011. View Article : Google Scholar
12	Sounni NE and Noel A: Targeting the tumor microenvironment for cancer therapy. Clin Chem. 59:85–93. 2013. View Article : Google Scholar
13	Lim YC, Kang HJ, Kim YS and Choi EC: All-trans-retinoic acid inhibits growth of head and neck cancer stem cells by suppression of Wnt/β-catenin pathway. Eur J Cancer. 48:3310–3318. 2012. View Article : Google Scholar : PubMed/NCBI
14	Li Y, Zhang T, Korkaya H, Liu S, Lee HF, Newman B, Yu Y, Clouthier SG, Schwartz SJ, Wicha MS and Sun D: Sulforaphane, a dietary component of broccoli/broccoli sprouts, inhibits breast cancer stem cells. Clin Cancer Res. 16:2580–2590. 2010. View Article : Google Scholar : PubMed/NCBI
15	Yang T and Rycaj K: Targeted therapy against cancer stem cells. Oncol Lett. 10:27–33. 2015.PubMed/NCBI
16	Leon G, MacDonagh L, Finn SP, Cuffe S and Barr MP: Cancer stem cells in drug resistant lung cancer: Targeting cell surface markers and signaling pathways. Pharmacol Ther. 158:71–90. 2016. View Article : Google Scholar
17	Lau WM, Teng E, Chong HS, Lopez KA, Tay AY, Salto-Tellez M, Shabbir A, So JB and Chan SL: CD44v8-10 is a cancer-specific marker for gastric cancer stem cells. Cancer Res. 74:2630–2641. 2014. View Article : Google Scholar : PubMed/NCBI
18	Mansour SF and Atwa MM: Clinicopathological significance of CD133 and ALDH1 cancer stem cell marker expression in invasive ductal breast carcinoma. Asian Pac J Cancer Prev. 16:7491–7496. 2015. View Article : Google Scholar : PubMed/NCBI
19	Nishikawa S, Konno M, Hamabe A, Hasegawa S, Kano Y, Fukusumi T, Satoh T, Takiguchi S, Mori M, Doki Y, et al: Surgically resected human tumors reveal the biological significance of the gastric cancer stem cell markers CD44 and CD26. Oncol Lett. 9:2361–2367. 2015.PubMed/NCBI
20	Tseng JY, Yang CY, Yang SH, Lin JK, Lin CH and Jiang JK: Circulating CD133(+)/ESA(+) cells in colorectal cancer patients. J Surg Res. 199:362–370. 2015. View Article : Google Scholar : PubMed/NCBI
21	Shitara K, Doi T, Nagano O, Imamura CK, Ozeki T, Ishii Y, Tsuchihashi K, Takahashi S, Nakajima TE, Hironaka S, et al: Dose-escalation study for the targeting of CD44v cancer stem cells by sulfasalazine in patients with advanced gastric cancer (EPOC1205). Gastric Cancer. 20:341–349. 2017. View Article : Google Scholar
22	Wang W, Dong LP, Zhang N and Zhao CH: Role of cancer stem cell marker CD44 in gastric cancer: A meta-analysis. Int J Clin Exp Med. 7:5059–5066. 2014.
23	Takahashi RU, Miyazaki H and Ochiya T: The role of microRNAs in the regulation of cancer stem cells. Front Genet. 4:2952014. View Article : Google Scholar : PubMed/NCBI
24	Liu C and Tang DG: MicroRNA regulation of cancer stem cells. Cancer Res. 71:5950–5954. 2011. View Article : Google Scholar : PubMed/NCBI
25	Ma C, Huang T, Ding YC, Yu W, Wang Q, Meng B and Luo SX: MicroRNA-200c overexpression inhibits chemoresistance, invasion and colony formation of human pancreatic cancer stem cells. Int J Clin Exp Pathol. 8:6533–6539. 2015.PubMed/NCBI
26	Yu D, Shin HS, Lee YS and Lee YC: miR-106b modulates cancer stem cell characteristics through TGF-β/Smad signaling in CD44-positive gastric cancer cells. Lab Invest. 94:1370–1381. 2014. View Article : Google Scholar : PubMed/NCBI
27	Wu Q, Yang Z, Wang F, Hu S, Yang L, Shi Y and Fan D: MiR-19b/20a/92a regulates the self-renewal and proliferation of gastric cancer stem cells. J Cell Sci. 126:4220–4229. 2013. View Article : Google Scholar : PubMed/NCBI
28	Takaishi S, Okumura T, Tu S, Wang SS, Shibata W, Vigneshwaran R, Gordon SA, Shimada Y and Wang TC: Identification of gastric cancer stem cells using the cell surface marker CD44. Stem Cells. 27:1006–1020. 2009. View Article : Google Scholar : PubMed/NCBI
29	Xu H, Tian Y, Yuan X, Wu H, Liu Q, Pestell RG and Wu K: The role of CD44 in epithelial-mesenchymal transition and cancer development. Onco Targets Ther. 8:3783–3792. 2015.
30	Li LC, Wang DL, Wu YZ, Nian WQ, Wu ZJ, Li Y, Ma HW and Shao JH: Gastric tumor-initiating CD44 (+) cells and epithelial-mesenchymal transition are inhibited by γ-secretase inhibitor DAPT. Oncol Lett. 10:3293–3299. 2015.
31	Zhang W and Li Y: miR-148a downregulates the expression of transforming growth factor-β2 and SMAD2 in gastric cancer. Int J Oncol. 48:1877–1885. 2016.PubMed/NCBI
32	Wang XH, Liu MN, Sun X, Xu CH, Liu J, Chen J, Xu RL and Li BX: TGF-beta 1 pathway affects the protein expression of many signaling pathways, markers of liver cancer stem cells, cytokeratins, and TERT in liver cancer HepG2 cells. Tumour Biol. 37:3675–3681. 2016. View Article : Google Scholar
33	Hoshino Y, Nishida J, Katsuno Y, Koinuma D, Aoki T, Kokudo N, Miyazono K and Ehata S: Smad4 decreases the population of pancreatic cancer-initiating cells through transcriptional repression of ALDH1A1. Am J Pathol. 185:1457–1470. 2015. View Article : Google Scholar : PubMed/NCBI
34	Zhao S, Ammanamanchi S, Brattain M, Cao L, Thangasamy A, Wang J and Freeman JW: Smad4-dependent TGF-beta signaling suppresses RON receptor tyrosine kinase-dependent motility and invasion of pancreatic cancer cells. J Biol Chem. 283:11293–11301. 2008. View Article : Google Scholar : PubMed/NCBI